Targeting human urinary metabolome by LC-MS/MS: a review.
Urine is a biological matrix that contains hundreds of metabolic end products which constitute the urinary metabolome. The development and advances on LC-MS/MS have revolutionized the analytical study of biomolecules by enabling their accurate identification and quantification in an unprecedented manner. Nowadays, LC-MS/MS is helping to unveil the complexity of urine metabolome, and the results obtained have multiple biomedical applications. This review focuses on the targeted LC-MS/MS analysis of the urine metabolome. In the first part, we describe general considerations (from sample collection to quantitation) required for a proper targeted metabolic analysis. In the second part, we address the urinary analysis and recent applications of four relevant families: amino acids, catecholamines, lipids and steroids.


The use of Taguchi's methods for the evaluation of industrial knife design.
Knives are used in industry, and particularly on meat packaging assembly lines, where a premium is placed on the speed of cutting. One consequence of this is that knife design and selection has been based on the speed of cutting as the most significant measure of a knife. In industry the cuts that knives make vary in difficulty with many factors such as thickness, contour, and temperature of the material being cut. These factors may change the speed with which these cuts are made. According to Taguchi's methods, a robust design is one that maintains high performance while remaining insensitive to changes in its environment. This means that a robust knife design would enable an employee to cut not only with as much speed as possible in one particular cutting scenario, but also with a consistently high speed in as many cutting scenarios as possible. An experiment with various knife designs was evaluated with Taguchi's method. Ten subjects cut outlined clay patterns in 15 min with each one of nine different knives. The nine different designs were generated from the following combinations; three grip sizes (44.4, 50.8 and 63.5 mm), three coupling angles between the grip and the blade (0, 45 and 90 degrees), three blade heights measured from the middle of the blade (10, 13 and 15 mm), and three blade lengths (85, 125 and 155 mm). Eight conditions of cutting material were used to generate 'noise'. They are combinations of temperature of the clay to be cut (from temperature (R) approximately 25 degrees C, frozen for 1 h (F) approximately 5 degrees C), thickness of the clay to be cut (15 and 25 mm), and orientation of the work surface (0 and 30 degrees). The result shows that the knife design of 63.5 mm grip size, 0 degree coupling angle, 13 mm blade height, and 85 mm blade length provides the highest number of cuts, the knife design of 50.8 mm grip size, 45 degrees coupling angle, 15 mm blade height, and 85 mm blade length gives the highest signal to noise ratio. The shorter blade (85 mm) is suggested by both designs, but the two designs are different in the other three variables. Further analysis of the noise variables shows that lowering clay temperature significantly reduces cutting speed. Although there is more than one design that can resolve changes in clay thickness and clay orientation, a 15 mm blade height is the only design insensitive to the clay temperature change.


Three-body wear of four types of light-activated composite resin veneering materials.
The in vitro wear resistance of four types of light-activated composite resins designed for crown and bridge veneers was determined by means of a three-body wear-testing device. After repeated cycles of loading, the wear values were measured with a profilometer, and surface textures were observed through a scanning electron microscope. All of the composite resin veneering materials examined in this investigation exhibited greater wear resistance than an unfilled poly(methyl methacrylate)-based material. The worn surface of veneering materials demonstrated outlines of fillers, indicating the difference in wear resistance between the resin matrix and the filler.


In vivo Imaging Technologies to Monitor the Immune System.
The past two decades have brought impressive advancements in immune modulation, particularly with the advent of both cancer immunotherapy and biologic therapeutics for inflammatory conditions. However, the dynamic nature of the immune response often complicates the assessment of therapeutic outcomes. Innovative imaging technologies are designed to bridge this gap and allow non-invasive visualization of immune cell presence and/or function in real time. A variety of anatomical and molecular imaging modalities have been applied for this purpose, with each option providing specific advantages and drawbacks. Anatomical methods including magnetic resonance imaging (MRI), computed tomography (CT), and ultrasound provide sharp tissue resolution, which can be further enhanced with contrast agents, including super paramagnetic ions (for MRI) or nanobubbles (for ultrasound). Conjugation of the contrast material to an antibody allows for specific targeting of a cell population or protein of interest. Protein platforms including antibodies, cytokines, and receptor ligands are also popular choices as molecular imaging agents for positron emission tomography (PET), single-photon emission computerized tomography (SPECT), scintigraphy, and optical imaging. These tracers are tagged with either a radioisotope or fluorescent molecule for detection of the target. During the design process for immune-monitoring imaging tracers, it is important to consider any potential downstream physiologic impact. Antibodies may deplete the target cell population, trigger or inhibit receptor signaling, or neutralize the normal function(s) of soluble proteins. Alternatively, the use of cytokines or other ligands as tracers may stimulate their respective signaling pathways, even in low concentrations. As in vivo immune imaging is still in its infancy, this review aims to describe the modalities and immunologic targets that have thus far been explored, with the goal of promoting and guiding the future development and application of novel imaging technologies.


The antibody binding site. Labelling of a specific antibody against the photo-precursor of an aryl nitrene.
The isolation of specific rabbit antibodies for the haptenic group 4-azido-2-nitrophenyl, is described. These antibodies bind 1.8-2.0mol of hapten [in-(4-azido-2-nitrophenyl)-l-lysine]/mol with an association constant of nearly 10(7)m(-1) at 4 degrees C. On photolysis of the antibody-hapten complex, resulting in the formation of an aryl nitrene at the binding site, hapten was covalently bound to the antibody, and the antibody binding site was blocked. The ratio of labelling of heavy- and light-chains was 2.5:1. Two small peptides were isolated from digests of labelled heavy-chain, indicating that some 13% of the label in the antibody was attached to cysteine-92 and to alanine-93. These residues are adjacent to the major hypervariable region in rabbit heavy-chain (residues 95-105).


[Application and therapeutic effect of frontal muscle fascia compound flap suspension for congenital blepharoptosis in children].
To investigate the application and therapeutic effect of frontal muscle fascia compound flap suspension for congenital blepharoptosis in children. From July 2010 to March 2012, 21 children (23 eyes) with congenital blepharoptosis were treated with frontal muscle fascia compound flap suspension. The therapeutic effect was observed and followed up. 17 patients were followed up for 3-6 months with relapse in one case. The ptosis was corrected in the other 1 cases. The frontal muscle fascia compound flap suspension is effective and practical with minimal morbidity and lower complication for the correction of congenital blepharoptosis.


[Cardiovascular malformations in Turner syndrome].
A literature review indicates that the prevalence of cardiovascular malformations in patients with Turner syndrome ranges from 17 to 47 percent. The aim of the study was to evaluate the cardiovascular system in patients with Turner syndrome, controlled by the Pediatric Endocrinology Clinic of the Medical University of Gdansk. The study included 55 females with Turner syndrome aged 3-20 years (mean age 9.6 years). The procedure included an interview, subject examination, chest X-rays and echocardiography. The echocardiography included M-mode measurements, two-dimensional evaluations from all standard planes, pulsed and continuous wave Doppler as well as color flow mapping. In selected cases, cardiologic examination embraced also cardiac catheterization, cineangiography and continuous 24-hour blood pressure measurement. Cardiovascular malformations were found in 18 out of 55 females (32.7%). Aortic coarctation was detected in 9 out of 18 patients (50%), bicuspid aortic valve in 33%, and aortic insufficiency in 22% of them. 78% of Turner subjects with cardiovascular malformations had monosomy (45,X). All females, except one, with aortic coarctation, had a 45,X karyotype. One aortic coarctation was diagnosed during prenatal examination. Before the last medical examination, 9 females had undergone a surgical treatment. In one case, the surgical treatment was preceded by a balloon angioplasty. Due to a recoarctation, one female was submitted to an additional balloon angioplasty. One female died during the post surgical period suffering from aortic wall injury. In four cases, despite successful surgeries, there still remains an arterial hypertension requiring pharmacological treatment. In 4 out of 18 cases (22%) isolated aortic valves defects were diagnosed, however, at present time they do not require any surgical treatment. In three cases, echocardiographic examination indicated a persistent left superior vena cava and aneurysm atrial septal defect with an intact septum. Due to a high prevalence of cardiovascular malformations, girls with Turner syndrome should be monitored by a pediatric cardiologist. In each case, female with Turner syndrome should be submitted to echocardiography. Also a careful control of the blood pressure is indicated.


Superior Mesenteric Artery Dissection After Lumbar Puncture.
We hereby present a case of iatrogenic dissection of the superior mesenteric artery dissection in a 63-year-old female undergoing a lumbar puncture (LP). She presented with severe diffused abdominal pain accompanied by lower back pain, nausea and vomiting a few hours after undergoing an LP due to ongoing headaches. Abdominal CT showed evidence of hemoperitoneum. She was then transferred to another facility and while in route received one unit of packed red blood cellsdue to drop in hemoglobin levels from 15 to 11 gm/dl. Physicians should consider the possibility of arterial variations and the level at which spinal tap is performed during interventions. Acute abdominal pain is a significant, common complaint that should be appropriately investigated.


[Therapeutic use of human leukocyte interferon in dermatologic disorders caused by herpes simplex virus. Multicenter study].
A multi-institutional double blind study was performed in 66 patients in order to evaluate the action of human leukocyte interferon, type alpha (IFN) in lesions produced by herpes simplex virus. Lesions were localized in genital area in 34 cases and in facial area in 32 of them. From the total, 38 patients were treated with 7,000 Ul/gm. of interferon ointment and 28 with carbowax 4,000 as placebo. The symptoms pain, ardor, itching and paresthesia were evaluated asking to patients. Erythema, vesicles, crust, scale adenopathy were objectivated. Signs and symptoms were controlled on days 3 and 7 of treatment. In 42 patients, lesions specimens were taken for virus isolation. In 23 of them, cytopathic effect was detected (54.76%). Results showed a rapid relief from pain (p less than 0.05), ardor (p less than 0.01) and paresthesia (p less than 0.001) and also accelerated healing of vesicles in patients treated with interferon. Tendency to reduce the total time of the disease in those patients treated with IFN was observed.


Brain gray and white matter abnormalities in preterm-born adolescents: A meta-analysis of voxel-based morphometry studies.
Studies using voxel-based morphometry report variable and inconsistent abnormalities of gray matter volume (GMV) and white matter volume (WMV) in brains of preterm-born adolescents (PBA). In such circumstances a meta-analysis can help identify the most prominent and consistent abnormalities. We identified 9 eligible studies by systematic search of the literature up to October 2017. We used Seed-based d Mapping to analyze GMV and WMV alterations between PBA and healthy controls. In the GMV meta-analysis, PBA compared to healthy controls showed: increased GMV in left cuneus cortex, left superior frontal gyrus, and right anterior cingulate cortex; decreased GMV in bilateral inferior temporal gyrus (ITG), left superior frontal gyrus, and right caudate nucleus. In the WMV meta-analysis, PBA showed: increased WMV in right fusiform gyrus and precuneus; decreased WMV in bilateral ITG, and right inferior frontal gyrus. In meta-regression analysis, the percentage of male PBA negatively correlated with decreased GMV of bilateral ITG. PBA show widespread GMV and WMV alterations in the default mode network, visual recognition network, and salience network. These changes may be causally relevant to socialization difficulties and cognitive impairments. The meta-regression results perhaps reveal the structural underpinning of the cognition-related sex differences in PBA.


Not Only Expansion: Proline Content and Density Also Induce Disordered Protein Conformation Compaction.
Intrinsically disordered proteins (IDPs) adopt a wide array of different conformations that can be constrained by the presence of proline residues, which are frequently found in IDPs. To assess the effects of proline, we designed a series of peptides that differ with respect to the number of prolines in the sequence and their organization. Using high-resolution atomistic molecular dynamics simulations, we found that accounting for whether the proline residues are clustered or isolated contributed significantly to explaining deviations in the experimentally-determined gyration radii of IDPs from the values expected based on the Flory scaling-law. By contrast, total proline content makes smaller contribution to explaining the effect of prolines on IDP conformation. Proline residues exhibit opposing effects depending on their organizational pattern in the IDP sequence. Clustered prolines (i.e., prolines with ≤2 intervening non-proline residues) result in expanded peptide conformations whereas isolated prolines (i.e., prolines with >2 intervening non-proline residues) impose compacted conformations. Clustered prolines were estimated to induce an expansion of ∼20% in IDP dimension (via formation of PPII structural elements) whereas isolated prolines were estimated to induce a compaction of ∼10% in IDP dimension (via the formation of backbone turns). This dual role of prolines provides a mechanism for conformational switching that does not rely on the kinetically much slower isomerization of cis proline to the trans form. Bioinformatic analysis demonstrates high populations of both isolated and clustered prolines and implementing them in coarse-grained molecular dynamics models illustrates that they improve the characterization of the conformational ensembles of IDPs.


[Ruptured Aneurysm of Bihemispheric Anterior Cerebral Artery A1-A2 junction:A Case Report].
We report a rare case with a ruptured aneurysm of the bihemispheric anterior cerebral artery(ACA), a rare anomaly of the ACA. A 50-year-old female experienced a sudden onset of severe posterior cervical pain. CT revealed diffuse subarachnoid hemorrhage(SAH)and intra cerebral hemorrhage of the left rectal gyrus. Cerebral angiography showed the left bihemispheric ACA and hypoplasia of the right ACA. A saccular aneurysm was indicated at the A1-A2 junction of the left bihemispheric ACA. A neck clipping was performed completely through the right transsylvian approach. There are several reports of an aneurysm with bihemispheric ACA, but all cases are distal ACA aneurysms. In this case, the aneurysm was proximal to the bihemispheric ACA and such aneurysms have not been reported.


Transforming vaccines supply chains in Nigeria.
Nigeria is the most populous country in Africa and in 2012 was suffering some of the lowest vaccination rates in the World. A combination of factors had resulted in a dysfunctional immunization cold chain and supply chain. Recognizing that the number of unimmunized children contributed to high levels of under-5-mortality, and that health MDGs would not be attained, Minister of State for Health Mohammed Pate launched a vaccines transformation project in 2013. In partnership with BMGF, GAVI, UNICEF, WHO, other donors and implementing partners the transformation journey has so far taken three years and achieved impressive results. It has though faced challenges along the way and with the financial burden of GAVI graduation facing Nigeria, the economic downturn and the decentralized funding of health services, the results are far from sustained. This paper documents the work undertaken at the Federal level and then highlights specific work undertaken in partnership with Lagos State Government. It identifies the importance of taking an end to end approach and looking at the root causes of weak system performance. The strategy combined simple innovations in how data was captured, recorded and used to drive decision making. It included a comprehensive and systematic approach to cold chain procurement, installation and maintenance with a shift to a culture of active cold chain maintenance that is performing with higher levels of uptime. It also included supply chain redesign at both the Federal and State level. Finally, it involved an institutional transformation at the National Primary Health Care Development Agency (NPHCDA) to establish a data driven Department of Logistics and Health Commodities (DLHC) to manage the many challenges in immunizing 7.5 million children annually. While results have been impressive, there have been many challenges and lessons learned on the way. As Nigeria gets ready for its graduation from GAVI, a robust agile performing cold chain and supply chain will be essential for the good health of Nigeria's children and its economy. The necessary transformation journey has only just begun.


Beta-hydoxy-beta-methylbutyrate supplementation affects Walker 256 tumor-bearing rats in a time-dependent manner.
Cancer cachexia affects intermediary metabolism with intense and general catabolism. Walker 256 tumor is a model injected either subcutaneously (Sc) or intraperitoneally (Ip), with different metabolic features. Beta-hydroxy beta-methylbutyrate (HMbeta) is a leucine metabolite with anti-catabolic properties, the aim of this study being to investigate its effects on metabolic parameters in both tumor models. Controls (subcutaneous control group (ScC) and intraperitoneal control group (IpC)) and supplemented animals (subcutaneous supplemented group (ScS) and intraperitoneal supplemented group (IpS)) showed these results. Protein Sc values were (47.8%) lower than Ip groups. Sc group fat content was (65.16%) higher than Ip groups. Liver glycogen value for Sc groups was (38.4%) higher than Ip groups. Muscle glycogen value for Sc groups were (2.75 times) higher than Ip groups. Corticosterone and insulin values were lower (44.53%) and higher (45.94%), respectively, in Sc when compared with Ip groups. Glucose and lactate values for ScS were the lowest (61.7% and 41.53%) compared to other groups. ScC glutamine value was the highest (40.8%) of all groups. Glutamate Sc values were (42.65%) lower than Ip groups. Sc groups showed greater survival time compared with Ip groups. ScS group showed 100% increase in survival time when compared with ScC. HMbeta supplementation can increase survival time and promotes metabolic changes in cancer-bearing animals, but it seems to work in a time-dependent manner.


Atypical Aeromonas salmonicida vapA type V and Vibrio spp. are predominant bacteria recovered from ballan wrasse Labrus bergylta in Scotland.
Healthy and/or moribund farmed and wild ballan wrasse Labrus bergylta (>0.5 to 900 g) were sampled from hatcheries (n = 2) and Atlantic salmon Salmo salar cage sites (n = 8) in Scotland between February 2016 and October 2018. Less than half of the sampled individuals (n = 43; 32.3%) had been vaccinated (autogenous polyvalent vaccine; dip and/or injection) against atypical furunculosis (type V and VI), while 20 (15.0%) fish were not vaccinated, and the rest (70 individuals, 52.7%) were of unknown vaccination status. Swab samples from skin lesions, gill, liver, spleen and kidney were inoculated onto a variety of bacteriological agar plates, and bacteriology identification and sequencing analysis was performed on significant bacterial colonies. Atypical Aeromonas salmonicida (aAs) vapA type V was the predominant bacterial species (70/215 bacterial isolates, 32.5% of bacterial samples; 43/117 positive individual fish, 36.8%) isolated in this survey followed by Vibrio species, which were the most geographically prevalent bacteria. Photobacterium indicum/profundum was also isolated from L. bergylta for the first time during this study. The collection of these bacterial isolates provides useful information for disease management. Identifying the aAs isolates involved in disease in ballan wrasse could provide vital information for improving/updating existing autogenous vaccines.


Distribution of a novel protein AgK114 expression in the normal tissues of adult mice: dual expression of AgK114 and growth hormone in anterior pituitary cells.
The novel antigen K114 (AgK114) has been previously identified in normal hamster skin, and its expression has been up-regulated accompanying tissue damages of the skin, although there is no information on its biological functions. To determine the physiological role of AgK114, we prepared anti-mouse AgK114 monoclonal antibody and studied its tissue distribution in healthy adult mice by immunocytochemistry. A widespread and unique expression of AgK114 peptide was found in the selected organs of various systems (hair follicle cells and sebaceous gland of skin, ciliated epithelial cells of trachea and bronchial tube, striated portion of submandibular gland, distal convoluted tubule cells of kidney, ciliated epithelial cells of oviduct, medulla of adrenal gland and anterior lobe of pituitary gland). Interestingly, dual expression of AgK114 peptide and growth hormone in somatotrophs was found in anterior lobe of pituitary gland by double immunocytochemistry. AgK114 peptide was expressed widely in many regionally well-defined cellular systems in various peripheral tissues, suggesting that AgK114 peptide may have some roles of physiological functions in these organs. The data from our current study have provided a rationale for further studies of functional roles of AgK114 peptide in a variety of organs or tissues under physiological conditions.


Catalytic and thermodynamic properties of β-glucosidases produced by Lichtheimia corymbifera and Byssochlamys spectabilis.
The objective of the present study was to optimize parameters for the cultivation of Lichtheimia corymbifera (mesophilic) and Byssochlamys spectabilis (thermophilic) for the production of β-glucosidases and to compare the catalytic and thermodynamic properties of the partially purified enzymes. The maximum amount of β-glucosidase produced by L. corymbifera was 39 U/g dry substrate (or 3.9 U/mL), and that by B. spectabilis was 77 U/g (or 7.7 U/mL). The optimum pH and temperature were 4.5 and 55 °C and 4.0 and 50 °C for the enzyme from L. corymbifera and B. spectabilis, respectively. β-Glucosidase produced by L. corymbifera was stable at pH 4.0-7.5, whereas the enzyme from B. spectabilis was stable at pH 4.0-6.0. Regarding the thermostability, β-glucosidase produced by B. spectabilis remained stable for 1 h at 50 °C, and that from L. corymbifera was active for 1 h at 45 °C. Determination of thermodynamic parameters confirmed the greater thermostability of the enzyme produced by the thermophilic fungus B. spectabilis, which showed higher values of ΔH, activation energy for denaturation (Ea), and half-life t(1/2). The enzymes were stable in the presence of ethanol and were competitively inhibited by glucose. These characteristics contribute to their use in the simultaneous saccharification and fermentation of vegetable biomass.


[Current views on acquired valvular heart disease and the results of treatment].
The article deals with the problems of diagnosis of acquired valvular defects (AVD) and indication to surgical treatment of AVD in accordance with international recommendations and modern knowledge of this issue. The following AVD diagnosis methods are described: heart auscultation, echocardiography, tests with regulated load, angiographic and radionuclide methods. Among pathological conditions, more attention is placed on stenoses, aortic, mitral and tricuspid valve insufficiency, IHD. Of importance is interpretation of the data obtained at diagnostic tests in terms of indications to cardiosurgical interventions, optimization of the terms and outcomes of the surgery. Statistics are provided on natural survival of patients and efficacy of surgical treatment.


A single-chain fragment against prostate specific membrane antigen as a tool to build theranostic reagents for prostate cancer.
Prostate carcinoma is the most common non-cutaneous cancer in developed countries and represents the second leading cause of death. Early stage androgen dependent prostate carcinoma responds well to conventional therapies, but relatively few treatment options exist for patients with hormone-refractory prostate cancer. One of the most suitable targets for antibody-mediated approaches is prostate specific membrane antigen (PSMA) which is a well known tumour associated antigen. PSMA is a type II integral cell-surface membrane protein that is not secreted, and its expression density and enzymatic activity are increased progressively in prostate cancer compared to normal prostate epithelium, thereby making PSMA an ideal target for monoclonal antibody imaging and therapy. To obtain a small protein that can better penetrate tissue, we have engineered a single-chain variable fragment (scFv) starting from the variable heavy and light domains of the murine anti-PSMA monoclonal antibody D2B. scFvD2B was analysed in vitro for activity, stability, internalisation ability and in vivo for targeting specificity. Maintenance of function and immunoreactivity as well as extremely high radiolabelling efficiency and radiochemical purity were demonstrated by in vitro assays and under different experimental conditions. Despite its monovalent binding, scFvD2B retained a good strength of binding and was able to internalise around 40% of bound antigen. In vivo we showed its ability to specifically target only PSMA expressing prostate cancer xenografts. Due to these advantageous properties, scFvD2B has the potential to become a good theranostic reagent for early detection and therapy of prostate cancers.


Changes in the hypothalamic serotonergic function may mediate the endocrine effects of melatonin.
The effects of a single injection of melatonin on serum thyroxine, corticosterone and prolactin levels, and the associated changes in the metabolism of serotonin (5-HT) in several hypothalamic regions of male rats kept under a 14-h light 10-h dark cycle (lights on at 08:00 h), are investigated. Melatonin (500 micrograms/kg, s.c.) or replacing saline was injected at 18:00 h, and 8 animals from treated and control groups were killed 1 h (19:00 h), 12 h (06:00 h) and 18 h (12:00 h) later. Melatonin significantly reduced serum thyroxine, corticosterone and prolactin levels within 1 h of treatment, indicating the existence of an acute inhibitory effect on hormonal secretion. Serum prolactin levels were increased 18 h after treatment, suggesting the implication of a different long-term regulatory mechanism. Injected melatonin induced several acute changes in the metabolism of 5-HT in the hypothalamus. Thus, a significant increase of 5-HT content in the preoptic area-anterior hypothalamic (POA-AH) and medial hypothalamic regions of melatonin-treated rats was observed within 1 h of treatment. The content of 5-hydroxyindoleacetic acid (5-HIAA) increased in medial hypothalamus, and the ratio 5-HIAA/5-HT decreased in POA-AH. The results show that melatonin acutely modifies the serotonergic activity in POA-AH and medial hypothalamus, and simultaneously inhibits thyroid, adrenal and pituitary hormonal secretion. The results are discussed suggesting that the hypothalamic serotonergic system could be an ideal mechanism underlying short-term melatonin effects on endocrine function.


Virus-specific DTH prevents contralateral retinitis following intracameral inoculation of HSV-2.
Following inoculation of HSV-1 (KOS strain) into the anterior chamber of one eye of a BALB/c mouse, the virus travels to the uninoculated contralateral eye and contributes to the devastating retinal necrosis which occurs in this eye 10-12 days p.i. In parallel experiments, HSV-2 (MS strain) was inoculated uniocularly via the anterior chamber route; animals were sacrificed at intervals and their eyes removed for histopathology and virus culture. Histopathological examination revealed that, in contrast to the retinal destruction observed in HSV-1 inoculated animals, the retina of the contralateral eye of HSV-2 injected animals was unaffected. Culture studies demonstrated that, similar to the spread of HSV-1 after anterior chamber inoculation, HSV-2 reached the contralateral eye in two temporally separate waves and also revealed that the amount of virus recovered from the uninoculated eye of HSV-2 infected animals was significantly less than that recovered from the uninoculated contralateral eye of HSV-1 injected mice. Further examination demonstrated that animals inoculated with HSV-2 via the anterior chamber route did not develop the suppression of virus-specific delayed hypersensitivity (DH) which is characteristic of HSV-1 induced anterior chamber associated immune deviation (ACAID). Taken together, the sparing of the contralateral retina and the ability to generate a virus-specific DH response suggest that DH assists in the preservation of the contralateral retina of animals inoculated with HSV-2 via the anterior chamber route by limiting virus replication and the amount of virus which reaches the uninoculated eye.


Immunohistochemical localization of steroidogenic enzymes in the corpus luteum and the placenta of the ribbon seal (Phoca fasciata) and steller sea lion (Eumetopias jubatus).
To study the luteal and placental function of pinnipeds, we analyzed the localization of steroidogenic enzymes (P450scc, 3 beta HSD and P450arom) in the corpus luteum and the placenta of ribbon seals (Phoca fasciata) and Steller sea lions (Eumetopias jubatus) immunohistochemically. P450scc and 3 beta HSD were present in all luteal cells of both species. Almost all of the luteal cells were immunostained for P450arom, while P450scc and 3 beta HSD were negatively immunostained in placentae and P450arom was present in the syncytiotrophoblast of placentae. These findings suggest that 1) corpora lutea of both species synthesize pregnenolone, progesterone and estrogen during the entire pregnancy period, and 2) like other terrestrial carnivores in the suborder Caniformia, placentae of both species do not have the capability for synthesizing progesterone in the latter half of active pregnancy period.


Requirement of a Functional Flavin Mononucleotide Prenyltransferase for the Activity of a Bacterial Decarboxylase in a Heterologous Muconic Acid Pathway in Saccharomyces cerevisiae.
Biotechnological production of cis,cis-muconic acid from renewable feedstocks is an environmentally sustainable alternative to conventional, petroleum-based methods. Even though a heterologous production pathway for cis,cis-muconic acid has already been established in the host organism Saccharomyces cerevisiae, the generation of industrially relevant amounts of cis,cis-muconic acid is hampered by the low activity of the bacterial protocatechuic acid (PCA) decarboxylase AroY isomeric subunit Ciso (AroY-Ciso), leading to secretion of large amounts of the intermediate PCA into the medium. In the present study, we show that the activity of AroY-Ciso in S. cerevisiae strongly depends on the strain background. We could demonstrate that the strain dependency is caused by the presence or absence of an intact genomic copy of PAD1, which encodes a mitochondrial enzyme responsible for the biosynthesis of a prenylated form of the cofactor flavin mononucleotide (prFMN). The inactivity of AroY-Ciso in strain CEN.PK2-1 could be overcome by plasmid-borne expression of Pad1 or its bacterial homologue AroY subunit B (AroY-B). Our data reveal that the two enzymes perform the same function in decarboxylation of PCA by AroY-Ciso, although coexpression of Pad1 led to higher decarboxylase activity. Conversely, AroY-B can replace Pad1 in its function in decarboxylation of phenylacrylic acids by ferulic acid decarboxylase Fdc1. Targeting of the majority of AroY-B to mitochondria by fusion to a heterologous mitochondrial targeting signal did not improve decarboxylase activity of AroY-Ciso, suggesting that mitochondrial localization has no major impact on cofactor biosynthesis.IMPORTANCE In Saccharomyces cerevisiae, the decarboxylation of protocatechuic acid (PCA) to catechol is the bottleneck reaction in the heterologous biosynthetic pathway for production of cis,cis-muconic acid, a valuable precursor for the production of bulk chemicals. In our work, we demonstrate the importance of the strain background for the activity of a bacterial PCA decarboxylase in S. cerevisiae Inactivity of the decarboxylase is due to a nonsense mutation in a gene encoding a mitochondrial enzyme involved in the biosynthesis of a cofactor required for decarboxylase function. Our study reveals functional interchangeability of Pad1 and a bacterial homologue, irrespective of their intracellular localization. Our results open up new possibilities to improve muconic acid production by engineering cofactor supply. Furthermore, the results have important implications for the choice of the production strain.


Comparative analysis of the Code of Professional Ethics in Bulgaria and the Hippocratic Oath, Declaration of Geneva and International Code of Medical Ethics.
In this paper I aim at making a comparative analysis of The Code of Professional Ethics in Bulgaria (CPEB), The Hippocratic Oath, The Declaration of Geneva, and The World Medical Association International Code of Medical Ethics. Two problems of special interest are explored: whether the leading principles of fundamental ethical codes are presented in CPEB and whether the code itself is relevant to the current medical professional and social situation in the country. The conclusion reached after a step-by-step analysis is that CPEB attempts to cover a wide range of principles and problems in medical practice and corresponds with the fundamental ethical codes. Although the code is criticized in some points, it could be very useful, provided that it is well publicized in the profession.


[Adult form of acid maltase deficiency presenting as progressive spinal muscular atrophy].
As far as could be elucidated, a sporadic manifestation of a slowly progressing muscular weakness and atrophy had commenced in a female patient symmetrically in the pelvic girdle and thigh region at the age of about 22 years. Only at the age of 55 years a precise neurological examination was done. On the basis of clinical and electromyographic data spinal muscular atrophy of the Kugelberg-Welander type was diagnosed. Muscular biopsy demonstrated severe far-progressed neurogenic muscular atrophy. In addition, increased storage of PAS-positive material was noticed. This was particularly true in the sparsely preserved, non-denervated muscular areas changed in the manner of an accompanying myopathy. Complementing electron microscopy and pathobiochemistry in the muscular biopsy showed an adult form of lack of acid maltase (Pompe's disease) aetiologically. This enzyme defect could also be demonstrated in the white cells of this patient.


Reproductive cycles of neotropical boid snakes evaluated by ultrasound.
Snakes have increasingly been bred as pets around the world. Few studies have addressed the reproduction of boid snakes, and no study has addressed their reproductive cycles in captivity. Thus, this paper describes the reproductive aspects of Brazilian boids in captivity. We used ultrasonography to characterize the reproductive cycle of four boid species in captivity in the Southern Hemisphere: the anaconda (Eunectes murinus), the red-tailed boa (Boa constrictor constrictor), the Amazon tree boa (Corallus hortulanus), and the rainbow boa (Epicrates cenchria). Nonvitellogenic follicles occurred from January to December in anaconda and red-tailed boa and for a shorter period from September to February in Amazon tree boa and from January to May in rainbow boa. Vitellogenesis occurred from late June to late March in E. murinus in year-round (12 months), from March to March in Amazon tree boa, from late September to late March in red-tailed boa, and from late March to late September in rainbow boa. Mating occurred from late March to late September in red-tailed boa and rainbow boa and from late September to late March in Amazon tree boa. No mating was observed in anacondas, but a female probably underwent parthenogenesis. Births occurred in July in anaconda and in March to July in Amazon tree boa and from December to March in red-tailed boa and rainbow boa. In males, increases in testicular size were associated with the mating season. Ultrasonography proved to be a safe and noninvasive technique to study the reproductive cycle of giant snakes in captivity.


Myotonic dystrophy type 1 with diabetes mellitus, mixed hypogonadism and adrenal insufficiency.
Myotonic dystrophy type 1 (DM1) is an autosomal dominant multisystem disease affecting muscles, the eyes and the endocrine organs. Diabetes mellitus and primary hypogonadism are endocrine manifestations typically seen in patients with DM1. Abnormalities of hypothalamic-pituitary-adrenal (HPA) axis have also been reported in some DM1 patients. We present a case of DM1 with a rare combination of multiple endocrinopathies; diabetes mellitus, a combined form of primary and secondary hypogonadism, and dysfunction of the HPA axis. In the present case, diabetes mellitus was characterized by severe insulin resistance with hyperinsulinemia. Glycemic control improved after modification of insulin sensitizers, such as metformin and pioglitazone. Hypogonadism was treated with testosterone replacement therapy. Notably, body composition analysis revealed increase in muscle mass and decrease in fat mass in our patient. This implies that manifestations of hypogonadism could be hidden by symptoms of myotonic dystrophy. Our patient had no symptoms associated with adrenal deficiency, so adrenal dysfunction was carefully followed up without hydrocortisone replacement therapy. In this report, we highlight the necessity for evaluation and treatment of multiple endocrinopathies in patients with DM1. DM1 patients could be affected by a variety of multiple endocrinopathies.Our patients with DM1 presented rare combinations of multiple endocrinopathies; diabetes mellitus, combined form of primary and secondary hypogonadism and dysfunction of HPA axis.Testosterone treatment of hypogonadism in patients with DM1 could improve body composition.The patients with DM1 should be assessed endocrine functions and treated depending on the degree of each endocrine dysfunction.


Mammalian genome evolution: new clues from comparisons of eutherians, marsupials and monotremes.
1. Comparisons of chromosomes and gene maps of different mammals are yielding a big picture of the evolution of mammalian genome form and function. It has been particularly instructive to compare gene arrangements on the sex chromosomes between the three major groups of mammals. Eutheria (so-called placental mammals). Metatheria (marsupials) and Prototheria (monotremes), which diverged 150 and 170 Myr BP respectively. 2. A region amounting to 3% of the haploid genome is located on the X chromosome in all three groups, implying that this region must have been part of the original X in a common ancestor. This region comprises the long arm of the human X. 3. A region represented by the short arm of the human X is common to the X in all eutherians, but is autosomal in marsupials and monotremes; thus it was not a part of the original X, and must have been acquired by the X early in the eutherian radiation. 4. This recently acquired region was probably translocated to a pseudoautosomal region shared by the eutherian X and Y. Thus it was originally paired and exempt from X chromosome inactivation; stepwise deletion of this region from the Y and recruitment of the newly unpaired region of the X into the inactivation system could account for some of the peculiarities of this region of the human X. 5. The sex-determining gene TDF must lie on the Y in all mammals in which the Y is male determining. The autosomal location of the candidate gene ZFY in marsupials and monotremes eliminates it from consideration. The recently described candidate gene SRY has yet to pass the "marsupial test".


Resilience, syndemic factors, and serosorting behaviors among HIV-positive and HIV-negative substance-using MSM.
Serosorting is commonly employed by MSM to reduce HIV risk. We hypothesize that MSM perceive serosorting to be effective, and that serosorting is predicted by resilience and inversely related to syndemic characteristics. Surveys included 504 substance-using MSM. Logistic regression models examined syndemic and resilience predictors of serosorting, separately by serostatus. For HIV-positive men, positive coping behaviors (P = .015) and coping self-efficacy (P = .014) predicted higher odds, and cognitive escape behaviors (P = .003) lower odds, of serosorting. For HIV-negative men, social engagement (P = .03) and coping self-efficacy (P = .01) predicted higher odds, and severe mental distress (P = .001), victimization history (P = .007) and cognitive escape behaviors (P = .006) lower odds, of serosorting. HIV-negative serosorters reported lower perceptions of risk for infection than non-serosorters (P < .000). Although high risk HIV-negative men may perceive serosorting to be effective, their high rates of UAI and partner change render this an ineffective risk reduction approach. Relevant public health messages are urgently needed.


Cloning, expression, purification, crystallization and preliminary X-ray analysis of NodS N-methyltransferase from Bradyrhizobium japonicum WM9.
The Nod factor (NF) is a rhizobial signal molecule that is involved in recognition of a legume host and the formation of root and stem nodules. Some unique enzymes are involved in the biosynthesis of NF, which is a variously but specifically substituted lipochitooligosaccharide. One of these enzymes is NodS, an N-methyltransferase that methylates end-deacetylated chitooligosaccharide substrates. In the methylation reaction, NodS uses S-adenosyl-L-methionine (SAM) as a methyl donor. To date, no structural information is available about NodS from any rhizobium. X-ray crystallographic studies of the NodS protein from Bradyrhizobium japonicum WM9, which infects the legumes lupin and serradella, have been undertaken. The nodS gene was cloned and the recombinant protein was expressed in Escherichia coli cells using natural amino acids and as an SeMet derivative. NodS without ligands was crystallized in the presence of PEG 3350 and MgCl(2). The protein was also crystallized in complex with S-adenosyl-L-homocysteine (SAH) in the presence of PEG 8000 and MgCl(2). SAH is produced from SAM as a byproduct of the methylation reaction. The crystals of apo NodS are tetragonal and diffracted X-rays to 2.42 A resolution. The NodS-SAH complex crystallizes in an orthorhombic space group and the crystals diffracted X-rays to 1.85 A resolution.


Neural cell adhesion molecule modulates mesenchymal stromal cell migration via activation of MAPK/ERK signaling.
Mesenchymal Stromal Cells (MSCs) represent promising tools for cellular therapy owing to their multipotentiality and ability to localize to injured, inflamed sites and tumor. Various approaches to manipulate expression of MSC surface markers, including adhesion molecules and chemokine receptors, have been explored to enhance homing of MSCs. Recently, Neural Cell Adhesion Molecule (NCAM) has been found to be expressed on MSCs yet its function remains largely elusive. Herein, we show that bone marrow-derived MSCs from NCAM deficient mice exhibit defective migratory ability and significantly impaired adipogenic and osteogenic differentiation potential. We further explore the mechanism governing NCAM mediated migration of MSCs by showing the interplay between NCAM and Fibroblast Growth Factor Receptor (FGFR) induces activation of MAPK/ERK signaling, thereby the migration of MSCs. In addition, re-expression of NCAM180, but not NCAM140, could restore the defective MAPK/ERK signaling thereby the migration of NCAM deficient MSCs. Finally, we demonstrate that NCAM180 expression level could be manipulated by pro-inflammatory cytokine Tumor Necrosis Factor (TNF)-α treatment. Overall, our data reveal the vital function of NCAM in MSCs migration and differentiation thus raising the possibility of manipulating NCAM expression to enhance homing and therapeutic potential of MSCs in cellular therapy.


Polymorphism and Molten Nitrate Salt-Assisted Single Crystal to Single Crystal Ion Exchange in the Cesium Ferrogermanate Zeotype: CsFeGeO4.
Two polymorphs of a new cesium ferrogermanate zeotype, CsFeGeO4, were synthesized using the molten CsCl-CsF flux growth approach at 900 °C. The orthorhombic polymorph, referred to as (1), crystallizes in the centrosymmetric nonpolar Pbcm space group. The compound exhibits a three-dimensional porous framework structure composed of disordered (Fe/Ge)O4 corner-sharing tetrahedra that generate large eight-sided channels running down the b-axis. These channels are occupied by Cs ions that provide charge balance to the anionic framework. Minor modifications in the reaction conditions lead to the synthesis of a monoclinic polymorph of CsFeGeO4, referred to as (2), crystallizing in the noncentrosymmetric polar space group P21 and exhibiting an identical framework structure to (1), albeit featuring ordered FeO4 and GeO4 tetrahedra. Solid state synthesis of CsFeGeO4 produces a polycrystalline mixture of (1) and (2), referred to as (6). Polarization-electric field (P-E) measurements of (6) indicate that the material is not ferroelectric. Powder second harmonic generation (SHG) measurements of (2) and (6) revealed them to be SHG active with intensities of 1.5 and 0.2 times that of α-SiO2, respectively. The temperature dependent magnetic susceptibility of (2) exhibits a downturn at T = 2.6 K, indicative of antiferromagnetic ordering. First-principles calculations in the form of density functional theory showed that (1) and (2) differ in stability by only 1.3 meV/atom, with (2) being the thermodynamically stabilized phase. Additional calculations for (1), using molten nitrate as reference, predicted the formation of energetically favorable phases, KFeGeO4 (3) and RbFeGeO4 (4). They were subsequently prepared via a molten nitrate salt bath treatment of (1) to replace Cs with K and Rb, affording (3) and (4) as single-crystal to single-crystal ion exchange products. Structure determination and property measurements for a pyroxene phase, CsFeGe2O6, referred to as (5), are also reported. This compound crystallized as a side product in the flux synthesis of CsFeGeO4.


Domestic violence protection measures.
In this article Richard Griffith, Senior Lecturer in Health Law at Swansea University, sets out the powers available to the police and court to protect people from domestic violence.


Clan genomics: From OMIM phenotypic traits to genes and biology.
Clinical characterization of a patient phenotype has been the quintessential approach for elucidating a differential diagnosis and a hypothesis to explore a potential clinical diagnosis. This has resulted in a language of medicine and a semantic ontology, with both specialty- and subspecialty-specific lexicons, that can be challenging to translate and interpret. There is no 'Rosetta Stone' of clinical medicine such as the genetic code that can assist translation and interpretation of the language of genetics. Nevertheless, the information content embodied within a clinical diagnosis can guide management, therapeutic intervention, and potentially prognostic outlook of disease enabling anticipatory guidance for patients and families. Clinical genomics is now established firmly in medical practice. The granularity and informative content of a personal genome is immense. Yet, we are limited in our utility of much of that personal genome information by the lack of functional characterization of the overwhelming majority of computationally annotated genes in the haploid human reference genome sequence. Whereas DNA and the genetic code have provided a 'Rosetta Stone' to translate genetic variant information, clinical medicine, and clinical genomics provide the context to understand human biology and disease. A path forward will integrate deep phenotyping, such as available in a clinical synopsis in the Online Mendelian Inheritance in Man (OMIM) entries, with personal genome analyses.


The First Report of CADASIL in Peru: Olfactory Dysfunction on Initial Presentation.
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) is a rare, heritable, small vessel vascular disease caused by mutations in the Notch3 gene that is characterized by migraines, subcortical vascular events, cognitive decline, and mood disturbances. However, many CADASIL cases present with unusual symptoms such as status epilepticus, a movement disorder, or sensory dysfunction. This study describes the clinical, genetic, and radiologic characteristics of a Peruvian family with CADASIL in which multiple family members presented with severe olfactory deficits. Seven members of the family have symptoms suggestive of CADASIL, with genetic testing revealing R133C mutations in the two patients who underwent genetic testing. Cognitive testing and olfactory identification testing (Smell Identification Test) were performed in three CADASIL patients revealing total anosmia in two tested patients and severe hyposmia in the other. Olfactory dysfunction has been associated with various neurologic and psychiatric conditions though few studies have linked it with neurovascular disorders such as CADASIL. This first reported case of CADASIL in Peru emphasizes that symptomatic olfactory dysfunction may be an unusual presentation of CADASIL and that olfactory dysfunction is important to evaluate in CADASIL patients.


Laser Microdissection.
Laser microdissection (LM) offers a relatively rapid and precise method of isolating and removing specified cells from complex tissues for subsequent analysis of their RNA, DNA, protein or metabolite content, thereby allowing assessment of the role of different cell types in the normal physiological or disease processes being studied. In this unit, protocols for the preparation of mammalian frozen tissues, fixed tissues, and cytologic specimens for LM, including tissue freezing, tissue processing and paraffin embedding, histologic sectioning, cell processing, hematoxylin and eosin staining, immunohistochemistry, and image-guided cell targeting are presented. Also provided are recipes for generating lysis buffers for the recovery of nucleic acids and proteins. The Commentary section addresses the types of specimens that can be utilized for LM and approaches to staining of specimens for cell visualization. Emphasis is placed on the preparation of tissue or cytologic specimens as this is critical to effective LM.


Antimicrobial polymers.
Better health is basic requirement of human being, but the rapid growth of harmful pathogens and their serious health effects pose a significant challenge to modern science. Infections by pathogenic microorganisms are of great concern in many fields such as medical devices, drugs, hospital surfaces/furniture, dental restoration, surgery equipment, health care products, and hygienic applications (e.g., water purification systems, textiles, food packaging and storage, major or domestic appliances etc.) Antimicrobial polymers are the materials having the capability to kill/inhibit the growth of microbes on their surface or surrounding environment. Recently, they gained considerable interest for both academic research and industry and were found to be better than their small molecular counterparts in terms of enhanced efficacy, reduced toxicity, minimized environmental problems, resistance, and prolonged lifetime. Hence, efforts have focused on the development of antimicrobial polymers with all desired characters for optimum activity. In this Review, an overview of different antimicrobial polymers, their mechanism of action, factors affecting antimicrobial activity, and application in various fields are given. Recent advances and the current clinical status of these polymers are also discussed.


Cells transiently expressing periostin are required for intramedullary intramembranous bone regeneration.
Intramembranous bone regeneration plays an important role in fixation of intramedullary implants used in joint replacement and dental implants used in tooth replacement. Despite widespread recognition of the importance of intramembranous bone regeneration in these clinical procedures, the underlying mechanisms have not been well explored. A previous study that examined transcriptomic profiles of regenerating bone from the marrow space showed that increased periostin gene expression preceded increases in several osteogenic genes. We therefore sought to determine the role of cells transiently expressing periostin in intramedullary intramembranous bone regeneration. We used a genetic mouse model that allows tamoxifen-inducible fluorescent labeling of periostin expressing cells. These mice underwent ablation of the bone marrow cavity through surgical disruption, a well-established intramembranous bone regeneration model. We found that in intact bones, fluorescently labeled cells were largely restricted to the periosteal surface of cortical bone and were absent in bone marrow. However, following surgical disruption of the bone marrow cavity, cells transiently expressing periostin were found within the regenerating tissue of the bone marrow compartment even though the cortical bone remained intact. The source of these cells is likely heterogenous, including cells occupying the periosteal surface as well as pericytes and endothelial cells within the marrow cavity. We also found that diphtheria toxin-mediated depletion of cells transiently expressing periostin at the time of surgery impaired intramembranous bone regeneration in mice. These data suggest a critical role of periostin expressing cells in intramedullary intramembranous bone regeneration and may lead to novel therapeutic interventions to accelerate or enhance implant fixation.


Predicting portal hypertension as assessed by acoustic radiation force impulse: correlations with the Doppler ultrasound.
The purpose of this study was to assess the value of acoustic radiation force impulse (ARFI) for evaluating portal hypertension by correlating the elasticity of liver cirrhosis, as measured by ARFI, with haemodynamic indices measured by Doppler ultrasound. We evaluated the data of a total of 154 prospectively enrolled patients who underwent both duplex Doppler ultrasound and ARFI imaging. The duplex Doppler ultrasound indices, including the mean portal vein velocity, splenic index (SI) and splenoportal index (SPI) were evaluated to determine the statistical correlation with shear wave velocity (SWV) of ARFI. We also analysed the differences in the correlations between the SI, SPI and SWV of the group who had varices. The correlations were assessed with Spearman's rank correlation coefficients. There was an increase of SWV in parallel with the increase of the SI (ρ=0.409, p<0.01) and SPI (ρ=0.451, p<0.01). In the patient group who did not have varices (n=74), the Doppler indices were found to be more correlated with the SWV (SI: ρ=0.447, SPI: ρ=0.552, p<0.01). However, the group with varices showed no correlation between the SWV and the Doppler parameters. SWV of ARFI was well correlated statistically with Doppler parameters, but is of limited value in predicting portal hypertension directly owing to great variability of Doppler parameters. By evaluating the correlation between ARFI and Doppler ultrasound, we suggest that the SWV might be a non-invasive supplementary tool for predicting portal hypertension.


Effects of clofibrate on some microsomal hydroxylations involved in the formation and metabolism of bile acids in rat liver.
1. The liver microsomal metabolism of [4-14C]cholesterol, endogenous cholesterol, 7 alpha-hydroxy-4-[6 beta-3H]cholesten-3-one, 5-beta-[7 beta-3H]cholestane-3 alpha, 7 alpha-diol and [3H]lithocholic acid was studdied in control and clofibrate (ethyl p-chlorophenoxyisobutyrate)-treated rats. 2. The extent of 7 alpha-hydroxylation of exogenous [414C]cholesterol and endogenous cholesterol, the latter determined with a mass fragmentographic technique, was the same in the two groups of rats. The extent of 12 alpha-hydroxylation of 7 alpha-hydroxy-4-cholesten-3-one and 5 beta-cholestane-3 alpha, 7 alpha-diol was increased by about 60 and 120% respectively by clofibrate treatment. The 26-hydroxylation of 5 beta-cholestane-3 alpha, 7 alpha-diol was not significantly affected by clofibrate. The 6 beta-hydroxylation of lithocholic acid was about 80% higher in the clofibrate-treated animals than in the controls. 3. The results are discussed in the context of present knowledge about the liver microsomal hydroxylating system and bile acid formation in patients with hypercholesterolaemia, treated with clofibrate.


Rationally designed foldameric adjuvants enhance antibiotic efficacy via promoting membrane hyperpolarization.
The negative membrane potential of bacterial cells influences crucial cellular processes. Inspired by the molecular scaffold of the antimicrobial peptide PGLa, we have developed antimicrobial foldamers with a computer-guided design strategy. The novel PGLa analogues induce sustained membrane hyperpolarization. When co-administered as an adjuvant, the resulting compounds - PGLb1 and PGLb2 - have substantially reduced the level of antibiotic resistance of multi-drug resistant Escherichia coli, Klebsiella pneumoniae and Shigella flexneri clinical isolates. The observed antibiotic potentiation was mediated by hyperpolarization of the bacterial membrane caused by the alteration of cellular ion transport. Specifically, PGLb1 and PGLb2 are selective ionophores that enhance the Goldman-Hodgkin-Katz potential across the bacterial membrane. These findings indicate that manipulating bacterial membrane electrophysiology could be a valuable tool to overcome antimicrobial resistance.


The Usage of Mock Oral Examinations for Program Improvement.
Mock oral examinations are often used to prepare residents for the American Board of Surgery certifying examination. Another potential use of these examinations is to identify programmatic weaknesses. Results from a multi-institutional mock oral examination were evaluated to determine if specific areas of weakness within each of the participating programs could be identified to facilitate program development. A mock oral examination was administered annually consisting of 3 examination rooms per resident with 3 cases in each room. Case categories included core general surgery and subspecialties and cases were changed yearly. Each case included facets of patient management from history and physical examination, and differential diagnosis to postoperative care and professional behaviors. General Surgery programs at 3 academic medical centers-Northwestern University, Rush University, and University of Illinois at Chicago. A total of 259 resident examinations of fourth- and fifth-year general surgery residents over a 7-year period. A total of 2331 individual resident cases were evaluated with an overall case pass rate of 50.2% across all 3 programs. The weakest case category for each program was different (A = vascular 40.0% pass, B = trauma 41.4% pass, and C = breast 30.0% pass). All programs scored above their mean in gastrointestinal and abdominal surgery and below their mean in vascular surgery. Within vascular surgery, the weakest facet of patient management also differed between programs (A = select tests 44.3% pass, B = complications 57.0% pass, and C = history and physical 55.4% pass). A standardized mock oral examination is able to identify topic areas of relative strength and weakness that differ between programs. These results can be used to define focused areas for improvement within training programs, guide rotation schedules, and improve didactic curricula.


Rapid Evidence Review of Bariatric Surgery in Super Obesity (BMI ≥ 50 kg/m2).
Despite accumulating evidence of the important health benefits of bariatric surgery in morbidly obese patients in general, bariatric surgery outcomes are less clear in higher-risk, high-priority populations of patients with BMI ≥ 50 kg/m2. To help the Department of Veterans Affairs (VA) Health Services Research & Development Service (HSR&D) develop a research agenda, we conducted a rapid evidence review to better understand bariatric surgery outcomes in adults with BMI ≥ 50 kg/m2. We searched MEDLINE®, the Cochrane Database of Systematic Reviews, the Cochrane Central Registry of Controlled Trials, and ClinicalTrials.gov through June 2016. We included trials and observational studies. We used pre-specified criteria to select studies, abstract data, and rate internal validity and strength of the evidence (PROSPERO registration number CRD42015025348). All decisions were completed by one reviewer and checked by another. Among 1892 citations, we included 23 studies in this rapid review. Compared with usual care, one large retrospective VA study provided limited evidence that bariatric surgery can lead to increased mortality in the first year, but decreased mortality long-term among super obese veterans. Studies that compared different bariatric surgical approaches suggested some differences in weight loss and complications. Laparoscopic gastric bypass generally resulted in greater short-term proportion of excess weight loss than did other procedures. Duodenal switch led to greater long-term weight loss than did gastric bypass, but with more complications. The published literature that separates the super obese is insufficient for determining the precise balance of benefits and harms of bariatric surgery in this high-risk subgroup. Future studies should evaluate a more complete set of key outcomes with longer follow-up in larger samples of more broadly representative adults.


Onset of Casimir-Polder retardation in a long-range molecular quantum state.
Two 4He atoms form a diatomic molecule with a significant vibrational wave function amplitude at interatomic separations R>100 Å, where the retardation switches the London R(-6) decay of the potential to the Casimir-Polder R(-7) form. It has been assumed that this effect of retardation on the long-range part of the potential is responsible for the 2 Å (4%) increase of the bond length <R> of 4He2. We show that <R> is, unexpectedly, insensitive to the potential at R>20 Å and its increase is due to quantum electrodynamics effects computed by us from expressions valid at short R--beyond the validity range of Casimir-Polder theory--that seamlessly extend this theory to distances relevant for properties of long molecules.


[Estimating nursing staff at a specialized liver transplantation unit: comparison between actual and anticipated situations].
The study was developed in a surgical clinic of a public-university hospital where, among the offered therapeutic modalities, they are accomplished, also, liver transplants. In that clinic they are interned patient that need caring of nursing of different complexities, from the intermediate from to the intensive ones. The objective was to compare the staff nursing existence in the unit for the installment of nursing attendance with the preconized for the Resolution COFEN no. 189/96. It's conclude that the existent staff nursing respects the preconized for this Resolution.


Effects of flavonoids on xanthine oxidation as well as on cytochrome c reduction by milk xanthine oxidase.
That flavonoids inhibit xanthine oxidase from cow milk was confirmed by measuring oxygen consumption with an oxygen electrode. In contrast, flavonoids did not inhibit glucose oxidase, another oxygen consuming enzyme. Among the flavonoids tested, quercetin, kaempferol, myricetin, chrysin, quercitrin, and morin were potent inhibitors of xanthine oxidase; their inhibition rates (%) were 80, 70, 69, 62, 59, and 51 at 100 microM (except chrysin at 50 microM), respectively. The xanthine oxidase-inhibiting activities of the flavonoids were not always well correlated with the suppressive activities of the flavonoids on cytochrome c reduction by a xanthine-xanthine oxidase system. The inhibition of xanthine oxidase by quercetin was not affected by cupric ion. The partition rates of the flavonoids between n-butanol and a buffer solution seemed to account for some of the inhibition.


Physical and genetic characterization of the glucitol operon in Escherichia coli.
The glucitol (gut) operon has been identified in the colony bank of Clark and Carbon (A. Sancar and W. D. Rupp, Proc. Natl. Acad. Sci. USA 76:3144-3148, 1979). We subcloned the gut operon by using paCYC184, pACYC177, and pBR322. The operon, which is encoded in a 3.3-kilobase nucleotide fragment, consists of the gutC, gutA, gutB, and gutD genes. The repressor of the gut operon seemed to be encoded in the region downstream from the operon. The gene products of the gut operon were identified by using maxicells. The apparent molecular weights of the glucitol-specific enzyme II (product of the gutA gene), enzyme III (product of the gutB gene), and glucitol-6-phosphate dehydrogenase (product of the gutD gene) were about 46,000, 13,500, and 27,000, respectively, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.


Longitudinal changes in tear cytokines and antimicrobial proteins in trachomatous disease.
Trachoma is a neglected tropical disease caused by ocular infection with Chlamydia trachomatis, where repeated infections and chronic inflammation can ultimately result in scarring, trichiasis and blindness. While scarring is thought to be mediated by a dysregulated immune response, the kinetics of cytokines and antimicrobial proteins in the tear film have not yet been characterised. Pooled tears from a Gambian cohort and Tanzanian cohort were semi-quantitatively screened using a Proteome Profiler Array to identify cytokines differentially regulated in disease. Based on this screen and previous literature, ten cytokines (CXCL1, IP-10, IFN-γ, IL-1β, IL-8, IL-10, IL-12 p40, IL-1RA, IL-1α and PDGF), lysozyme and lactoferrin were assayed in the Tanzanian cohort by multiplex cytokine assay and ELISA. Finally, CXCL1, IP-10, IL-8, lysozyme and lactoferrin were longitudinally profiled in the Gambian cohort by multiplex cytokine assay and ELISA. In the Tanzanian cohort, IL-8 was significantly increased in those with clinically inapparent infection (p = 0.0086). Lysozyme, IL-10 and chemokines CXCL1 and IL-8 were increased in scarring (p = 0.016, 0.046, 0.016, and 0.037). CXCL1, IP-10, IL-8, lysozyme and lactoferrin were longitudinally profiled over the course of infection in a Gambian cohort study, with evidence of an inflammatory response both before, during and after detectable infection. CXCL1, IL-8 and IP-10 were higher in the second infection episode relative to the first (p = 0.0012, 0.044, and 0.04). These findings suggest that the ocular immune system responds prior to and continues to respond after detectable C. trachomatis infection, possibly due to a positive feedback loop inducing immune activation. Levels of CXC chemokines in successive infection episodes were increased, which may offer an explanation as to why repeated infections are a risk factor for scarring.


The nurse role in parent-infant bonding. Overview, assessment, intervention.
The recognition of appropriate and inappropriate attachment behavior is vital to maternal health and self-esteem and effective infant growth and development. An overview of parent-infant bonding is presented, followed by a discussion of the principles and methods of assessment and intervention.


Poliovirus infection and expression of the poliovirus protein 2B provoke the disassembly of the Golgi complex, the organelle target for the antipoliovirus drug Ro-090179.
Infection of Vero cells with poliovirus results in complete disassembly of the Golgi complex. Milestones of the process of disassembly are the release to the cytosol of the beta-COP bound to Golgi membranes, the disruption of the cis-Golgi network into fragments scattered throughout the cytoplasm, and the disassembly of the stacked cisternae by a process mediated by long tubular structures. Transient expression of the viral protein 2B in COS-7 cells also causes the disassembly of the Golgi complex by a process preceded by the accumulation of the protein in the Golgi area. Vero cells infected for 3 h show no recognizable Golgi complexes at the ultrastructural level and display an enormously swollen endoplasmic reticulum (ER) with extensive areas of its surface heavily coated. Ro-090179 (Ro), a flavonoid isolated from the herb Agastache rugosa, provokes the specific swelling and disruption of the Golgi complex and strongly inhibits poliovirus infection. Ro provokes the swelling and the disruption of the stacked cisternae and trans-Golgi elements without affecting the cis-most Golgi cisternae much. Moreover, Ro inhibits the fusion of the Golgi complex with the ER in cells treated with brefeldin A and provokes the accumulation of the intermediate compartment membrane protein p58 into ERD2-positive Golgi elements but has no effect on the anterograde transport involved in protein secretion. Our results indicate that the secretory pathway and specifically the Golgi complex are preferential targets of poliovirus.


[Limited effect of gonadotrophin-releasing hormone analogues for patients with endometriosis].
A Cochrane review concluded that gonadotrophin-releasing hormone analogues (GnRHas) are more effective at relieving endometriosis-associated pain than no treatment/placebo, while there was not found any difference in pain relief between GnRHas and danazol or between GnRHas and intrauterine levonorgestrel. A high frequency of hypoestrogenic side effects was found for GnRHas, since none of the studies included add-back therapy. This review confirmed that GnRHas can be used for endometriosis therapy, but first choice of medical treatment should be oral contraceptives or intrauterine levonorgestrel.


A delay discounting task produces a greater likelihood of waiting than a deferred gratification task.
A first-person-shooter video game was adapted for the study of choice between smaller sooner and larger later outcomes to compare the behavioral patterns produced by deferred gratification (DG) and delay discounting (DD) tasks. Participants played a game in which they could either fire their weapon sooner and do a small amount of damage or wait a few seconds to fire their weapon and do a larger amount of damage. For the DD task, a failure to fire within one second committed the player to waiting for the larger later outcome thus removing the opportunity to defect during the delay that is present in the DG task. The incentive structure changed multiple times during game play so that at times the optimal decision was to choose the smaller sooner outcome whereas at other times the optimal decision was to wait for the larger later outcome. Players assigned to the DD task showed a greater tendency to wait and lower sensitivity to the changing incentives.


The α-cell in diabetes mellitus.
Findings from the past 10 years have placed the glucagon-secreting pancreatic α-cell centre stage in the development of diabetes mellitus, a disease affecting almost one in every ten adults worldwide. Glucagon secretion is reduced in patients with type 1 diabetes mellitus, increasing the risk of insulin-induced hypoglycaemia, but is enhanced in type 2 diabetes mellitus, exacerbating the effects of diminished insulin release and action on blood levels of glucose. A better understanding of the mechanisms underlying these changes is therefore an important goal. RNA sequencing reveals that, despite their opposing roles in the control of blood levels of glucose, α-cells and β-cells have remarkably similar patterns of gene expression. This similarity might explain the fairly facile interconversion between these cells and the ability of the α-cell compartment to serve as a source of new β-cells in models of extreme β-cell loss that mimic type 1 diabetes mellitus. Emerging data suggest that GABA might facilitate this interconversion, whereas the amino acid glutamine serves as a liver-derived factor to promote α-cell replication and maintenance of α-cell mass. Here, we survey these developments and their therapeutic implications for patients with diabetes mellitus.


Effects of Escherichia coli and Porphyromonas gingivalis lipopolysaccharide on pregnancy outcome in the golden hamster.
This report describes the effects of two gram-negative bacterial endotoxin (lipopolysaccharide [LPS]) preparations on hamster pregnancy outcome variables. Single intravenous challenges with Escherichia coli and Porphyromonas gingivalis LPS on day 8 of pregnancy produced dose-dependent effects on fetal weight malformation and fetal resorption with E. coli LPS having potent embryolethal effects. Premating maternal exposure to P. gingivalis produced embryolethal effects similar to those of E. coli. These data suggest that maternal exposure to P. gingivalis LPS prior to and during pregnancy can induce deleterious effects on the developing fetus.


Left ventricular diastolic capacity in man.
Plasma volume expansion with 500 ml of low-molecular-weight dextran was used in 27 patients (nine normal subjects, 13 patients with ischemic heart disease, four with aortic stenosis and one with cardiomyopathy) to increase left ventricular end-diastolic pressure (LVEDP) from a control value of 12.4 +/- 7.0 mm Hg (mean +/- SD) to 23.3 +/- 7.0 mm Hg and end-diastolic volume (EDV) from 84.0 +/- 23.8 ml/m2 to 97.6 +/- 22.9 ml/m2. EDV-LVEDP curves constructed for 12 patients from multiple angiograms at progressively increasing LVEDPs during plasma volume expansion showed an initial part where EDV increased in parallel with LVEDP and a final steep or perpendicular part where EDV increased minimally or not at all as LVEDP exceeded 20 mm Hg. Exponential equations were used to fit diastolic volume-pressure data obtained with catheter-tip manometers in seven patients: the exponential constant, k, was 0.012-0.044 ml-1 and was inversely related to EDV (Spearman's rank correlation coefficient = -1). For comparable EDV, there were no differences in k values between normal subjects and patients with a variety of heart diseases.


Diffusion-weighted magnetic resonance imaging in prostate cancer patients on active surveillance one year after diagnosis and before repeat biopsy.
The aim of this study was to evaluate prospectively whether diffusion-weighted magnetic resonance imaging (DW-MRI), interpreted in a routine clinical setting, can serve as a diagnostic and prognostic tool for prostate carcinoma patients on active surveillance (AS). Eighty men enrolled in the Finnish arm of the PRIAS (Prostate Cancer Research International: Active Surveillance) study were followed for at least 1 year and had DW-MRI scans taken in addition to repeat biopsy. Spearman's correlations were analysed between tumour appearance on DW-MRI and clinical variables [age, prostate-specific antigen (PSA), free PSA, PSA doubling time, prostate volume, percentage of cancer at diagnostic biopsy]. The Pearson chi-squared test clarified associations between outcome factors (number of positive cores and Gleason score on repeat biopsy, treatment change) and DW-MRI results. Assumed predictors of deferred radical treatment were examined with logistic regression analysis. Accuracy of tumour localization by DW-MRI compared to repeat biopsy findings was analysed by the chi-squared test. DW-MRI revealed an anatomical lesion suggestive of cancer in 40 patients (50%). MRI positivity showed no significant correlation with clinical variables. No associations existed between tumour appearance on DW-MRI and biopsy findings or discontinuation of AS. The only variable predicting treatment change was higher PSA at discontinuation (p = 0.002). Appearance of tumour, either on T2-weighted MRI (p = 0.273) or on apparent diffusion coefficient maps (p = 0.691), was not a significant predictor of treatment change. Localized low-grade prostate cancer is challenging to visualize in DW-MRI, and this imaging technique provides no additional prognostic benefit compared to PSA and repeat biopsies.


Fatal cerebral coenurosis in a cat.
A 6-year-old cat that was laterally recumbent and panting was evaluated because of a 10-day history of progressive neurologic abnormalities. Despite aggressive treatment, the cat died on the day of admission. At necropsy, a 1.5-cm-diameter, fluid-filled cyst was found in the white matter of the left cerebrum. The cyst was identified as a coenurus of Taenia serialis, on the basis of the cyst wall, distribution of scolices, and the shape and dimensions of rostellar hooks. Scolices were found in varying stages of ontogeny, ranging from undifferentiated to nearly mature. Taenia serialis has a canid-lagomorph life cycle, with cats being accidental intermediate hosts. However, the potential exists for rare zoonotic transmission and subsequent serious disease in human beings and other accidental intermediate hosts.


[Long-term results of treatment of patients with large or giant intracranial aneurysms of internal carotid artery].
At Burdenko Neurosurgical Institute annually arrives 50-60 patients with large and giant aneurysms of the brain, which is nearly 15% of all patients with aneurysms. About half of them--patients with paraclinoid aneurysms, which are successfully treated by cliping using techniques of intravascular blood aspiration (IVBA) or proximal control of blood flow in the neck. This paper analyzes the long-term outcomes of patients who were operated for large and giant aneurysms of the internal carotid artery in the last 15 years. Catamnesis was collected from 93 (71.5%) of 130 patient. Mean follow-up was 65.2 +/- 36.0 months. Mean age of patients at the time of surgery was 45.55 +/- 7.5 years and at the time of catamnesis evaluation 50.4 +/- 10.2 years. Men to women ratio was 18:75. Patients with multiple aneurysms was 22.6% (21 patients). In most cases--67 (72%) patients--aneurysms were treated by cliping of aneurysms using IVBA, in 14 (15%) cases--using proximal control of blood flow in the neck, in 5 (5.4%) cases--reinforced with gauze and glue, in 6 (6.5%) patient--by aneurysm trapping, in one case the aneurysm was not cliped. Radical surgery performed in 87 (93.5%) patients. Catamnesis analysis of survived patients showed that 55.8% of them are complaining of headaches, which they associate with the operation. 39 (43.3%) patients noted persistent elements of asthenia, 4 (4.6%) had sleep disorders, 5 (5.8%) suffered of epileptic seizures and 1 (1.10%) of depression. Korsakoff's syndrome, developed after surgery in 2 (2.3%) patients remains. Only 38.4% of patients after surgery continue to work. Favorable outcome on the Glasgow outcome scale (4-5) was obtained in 81 (87.1%) patients, poor (2-3) in 5 (5.4%) patients. Mortality rate was 7.5%. Severe oculomotor disturbances have a slight tendency to recovery over time.


Intercellular communication mediates the bystander effect during herpes simplex thymidine kinase/ganciclovir-based gene therapy of human gastrointestinal tumor cells.
A bystander effect is described when nontransduced or genetically unmodified cells are killed during death of genetically modified tumor cells transduced with a suicide gene. The "bystander effect" greatly enhances the efficacy of the herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) gene therapy approach for cancer. The mechanism of the bystander effect is controversial. In this study, we examined the role of intercellular gap junction communication (GJIC) for the bystander effect in human gastrointestinal tumor cells. Our results show that the extent of the bystander effect varied amongst the tumor cell lines; pancreatic cancer cells BXPC-3 exhibited excellent bystander effects in vitro and in vivo studies whereas other gastrointestinal tumor cell lines such as pancreatic cancer cells MIAPACA-2, and colon cancer cells HT-29 showed poor bystander effects. Bystander effects were only found in the presence of cell-to-cell contact. The extent of the bystander effect was independent of the level of HSV-TK activity in the transduced tumor cells and was correlated with GJIC as demonstrated by an in vitro dye-transfer assay. Expression of the mRNA levels of gap junction protein connexin 43 was 8- to 26-fold or greater and connexin 26 gene expression was 2- to 229-fold greater in BXPC-3 cells compared to HT-29, MIAPACA-2, and PANC3 cells. Our results suggest that intercellular communication is essential for the bystander effect. The correlation between GJIC and the extent of the bystander effect suggest a role for GJIC in mediating the bystander effect. Analysis of tumors for GJIC or expression of gap junction proteins may identify the subset of patients suitable for gene therapy with the HSV-TK/GCV approach.


Unusual radiological appearance of a faecaloma.
Faecaloma is a hardened, relatively immobile, tumour-like mass of accumulated faeces, usually found in the rectosigmoid region. Rarely, a faecaloma may be observed in other locations along the colon. It is a rare condition but has important complications, including ulceration, perforation and obstruction. The diagnosis is important not only to avoid confusion with neoplasm but also to alert the physician to the possibility of complications. Here we present a case of faecaloma, including the clinical and radiological difficulties in the diagnosis.


The relationship of androgen to the thyrotropin and prolactin responses to thyrotropin-releasing hormone in hypogonadal and normal men.
Fluoxymesterone, an androgen not converted to estrogen, caused a significant decrease in the TSH response to TRH in 11 men with primary hypogonadism [maximum change in TSH: before treatment, 11.3 +/- microU/ml (mean +/- SE); 8.9 +/- 1.0 after 2 weeks (P less than 0.001); 8.2 +/- 1.1 after 6 weeks (P less than 0.01)]. There was a significant fall in serum T4-binding globulin (TBG) (measured directly by RIA) without a change in the free T4 or free T3 index. Fluoxymesterone had no effect on the PRL response to TRH in hypogonadal men. The results suggest that 1) androgen per se is at least partly responsible for the lower TSH response to TRH in men compared to women and 2) androgen is not a cause of the lower PRL response to TRH in men.


Prevalence and determinants of symptoms of common mental disorders in retired professional Rugby Union players.
The primary aim of this study was to determine the prevalence of symptoms of common mental disorders (CMD) (distress, anxiety/depression, sleeping disturbance, adverse nutrition behaviour, adverse alcohol behaviour and smoking) among retired professional Rugby Union players. The secondary aim was to explore the associations between stressors (life events, Rugby Union career dissatisfaction) and the health conditions under study. Therefore, cross-sectional analyses were conducted on baseline questionnaires from an ongoing prospective cohort study of retired professional Rugby Union players. An electronic questionnaire was established using validated questionnaires to assess symptoms of CMD and stressors. The electronic questionnaire was subsequently distributed to retired players by the national Rugby Union players' associations in France, Ireland and South Africa. Among 295 retired professional Rugby Union players (mean age of 38 years), prevalence rates were 25% for distress, 28% for anxiety/depression, 29% for sleeping disturbance, 62% for adverse nutrition behaviour, 15% for smoking and 24% for adverse alcohol behaviour. A higher number of life events were associated with distress (OR = 1.2; 95% CI 1.1-1.4), anxiety/depression (OR = 1.6; 95% CI 1.2-2.1), sleeping disturbance (OR = 1.6; 95% CI 1.2-2.1) and adverse nutrition behaviour (OR = 1.8; 95% CI 1.3-2.5). A higher level of dissatisfaction of the player's Rugby Union career was associated with distress (OR = 0.9; 95% CI 0.8-1.0), sleeping disturbance (OR = 0.9; 95% CI 0.9-1.0), smoking (OR = 0.9; 95% CI 0.9-1.0) and adverse nutrition behaviour (OR = 0.9; 95% CI 0.8-0.9). In conclusion, our study suggests that prevalence of symptoms of CMD is high among retired professional Rugby Union players, being associated with both a higher number of life events and a higher level of Rugby Union career dissatisfaction.


Analysis and simulation of gain control and precision in crayfish visual interneurons.
Impulse trains in sustaining and dimming fibers of crayfish optic lobe (in situ) were elicited with sinusoidal extrinsic current and sine-wave illumination. Extrinsic currents and currents derived from postsynaptic potentials (PSPs) were used to compute the time course of the spike train with an adaptive integrate-and-fire model. The neurons exhibit variations in gain and spike timing precision related to the frequency of stimulation. These phenomena are influenced by spike-frequency adaptation and nonlinearities in the PSP. Dimming fibers exhibit relatively strong spike-frequency adaptation and an associated increase in gain with the frequency of sinusoidal extrinsic current and sine-wave illumination. The dimming fiber IPSP promotes spike train rectification, and rectification contributes to spike timing precision. Sustaining fibers exhibit weaker spike-frequency adaptation and the gain of the current-elicited response is less sensitive to stimulus frequency. The sustaining fiber excitatory PSP, however, exhibits a strong frequency-dependent nonlinearity that influences the frequency response. Spike timing precision is a function of stimulus frequency in all cells and it is enhanced by rectification of the discharge and/or resonance. In rectified responses the jitter in spike times is closely related to the variance in the times the membrane potential reaches spike threshold. These gain and spike timing results are well approximated by the simulated responses. Because the nonlinearity of the sustaining fiber PSP entails a high rate of depolarization, the PSP can increase the precision of spike timing by 10- to 100-fold compared with the response to pure sine-wave stimuli. This enhanced precision has implications for crayfish oculomotor reflexes that are driven by sustaining fibers and highly sensitive to impulse timing during transient excitation.


Triplicated thumbs: a rarity?
Triplication of the thumb is supposed to be a rare condition and a complex form of radial polydactyly. However, we encountered an unusually high number of triplicated thumbs at our unit. Is triplication of the thumb indeed a rare condition? In our study, 121 patients with radial polydactyly were recorded between 1933 and 2005. In nine patients of this group, triplicated thumbs, either unilateral or bilateral, were identified. This complex type of radial polydactyly occurs in various forms. A total of 11 triplicated thumbs were found, and all cases were combined with triphalangeal components. Only one of these 11 triplicated thumbs could be classified according to currently used classifications. In all cases, aberrant rays were excised, thumb length and alignment restored by osteotomies, joints were stabilised, tendons reinserted and nails and nail walls corrected if necessary. Also, in all cases, a correction of triphalangeal components was carried out. Triplications are a rarity, and have only been presented in single case reports. We found that triplicated thumbs are not so rare. The likely explanation for this is the identification of a genetic isolate (a deformity located at chromosome 7q36) with radial polydactyly and triphalangeal thumbs in the southwest region of the Netherlands. Treatment for the presented triplicated thumbs was based on the same general principles as for less complex forms of radial polydactyly (i.e. to assemble useful elements of the separate [partial] thumbs to reconstruct one functioning, stable thumb).


A meta-analysis of stroke risk following herpes zoster infection.
The incidence of herpes zoster (HZ) is increasing and poses a significant health concern to aging populations. Several studies suggest an increased risk of stroke following zoster infection, but the results are conflicting. We conducted a systematic review and meta-analysis to determine if stroke risk is increased following HZ infection. A search of MEDLINE, EMBASE, Google scholar, Web of Science, CAB Direct, Cumulative Index to Nursing and Allied Health Literature, and Evidence Based Medicine Reviews was conducted for observational studies of adults with HZ infection that examined stroke and TIA risk from January 1, 1966 to May 31, 2016. Adjusted relative risks reported for similar follow-up durations were pooled across studies separately using random-effects inverse variance models. Data were pooled from nine studies. Relative risk for stroke after zoster was 1.78 (95% CI 1.70-1.88) for the first month following herpes zoster, dropping progressively to 1.43 (95% CI 1.38-1.47) after 3 months, to 1.20 (95% CI 1.14-1.26) after 1 year. We found that stroke risk increases by a larger margin during the first month after a herpes zoster ophthalmicus episode: relative risk 2.05 (95% CI 1.82-2.31). The risk remains elevated one year after the acute episode. Herpes zoster is an established risk factor for increasing the risk of stroke, especially shortly after infection. Vaccination should be encouraged in patients at high risk of cardiovascular disease.


Ectopic testis: a rare case.
Congenital undescending testis is a common anomaly of testis, but we had a rare case of ectopic testis. A 15-month-old infant was operated emergently because of left incarcerate inguinal hernia. Intraoperative exploration of hernial sac revealed two ectopic testes with one spermatic cord proximally but in the middle divided to two spermatic cords in a 8 shape. There was an important point about vas deferens as it was single proximal to the chord, but divided into two in the middle of the chord. Vessels showed a similar condition about. We released both testes and brought down both of them into scrotum. This is a rare case of ectopic testis transectopia with partially common vas and vessels.


On the level: IRGM gene function is all about expression.
Crohn disease is a complex, multigenic, chronic inflammatory bowel disease of uncertain etiology. Recent advances in genetics, including high-throughput single-nucleotide polymorphism typing platforms and deep sequencing technologies have begun to shed light upon disease predisposition and pathogenesis. Autophagy is emerging as a key player in both innate and adaptive immunity, as well as tissue homeostasis and development in the gut. Here we describe our recent studies into the Crohn disease-associated Immunity-Related GTPase family, M (IRGM) gene and our discovery of a large risk-conferring upstream deletion. We discuss the effects of this deletion upon expression levels of IRGM alleles and how tissue-specific expression might be affected by the promoter polymorphism. In addition, we comment upon the potential roles of IRGM in autophagy of intracellular pathogens, and the challenges ahead for further elucidating IRGM function.


Quantitation of spin probe-detectable oxidants in cells using electron paramagnetic resonance spectroscopy: To probe or to trap?
Electron Paramagnetic Resonance (EPR) spectroscopy coupled with spin traps/probes enables quantitative determination of reactive nitrogen and oxygen species (RNOS). Even with numerous studies using spin probes, the methodology has not been rigorously investigated. The autoxidation of spin probes has been commonly overlooked. Using the spin probe 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH), the present study has tested the effects of metal chelators, temperature, and oxygen content on the autoxidation of spin probes, where an optimized condition is refined for cell studies. The apparent rate of CMH autoxidation under this condition is 7.01 ± 1.60 nM/min, indicating low sensitivity and great variation of the CMH method and that CMH autoxidation rate should be subtracted from the generation rate of CMH-detectable oxidants (simplified as oxidants below) in samples. Oxidants in RAW264.7 cells are detected at an initial rate of 4.0 ± 0.7 pmol/min/106 cells, which is not considered as the rate of basal oxidants generation because the same method has failed to detect oxidant generation from the stimulation of phorbol-12-mysirate-13-acetate (PMA, 0.1 nmol/106 cells) in cells (2.5 ± 0.9 for PMA vs. 2.1 ± 1.5 pmol/min/106 cells for dimethyl sulfoxide (DMSO)-treated cells). In contrast, the spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), which exhibits minimal autoxidation, reveals differences between PMA and DMSO treatment (0.26 ± 0.09 vs. -0.06 ± 0.12 pmol/min/106 cells), which challenges previous claims that spin probes are more sensitive than spin traps. We have also found that low temperature EPR measurements of frozen samples of CMH autoxidation provide lower signal intensity and greater variation compared to RT measurements of fresh samples. The current study establishes an example for method development of RNOS detection, where experimental details are rigorously considered and tested, and raises questions on the applications of spin probes and spin traps.


Scaling of the density of state of the weighted Laplacian in the presence of fractal boundaries.
Spectral properties of the weighted Laplace operator in the presence of fractal boundaries are numerically investigated for both Neumann and Dirichlet boundary conditions. This corresponds to the characterization of heat and mass transport in microchannels with irregular and rough surfaces induced by the microfabrication process. The axial velocity field with no-slip boundary conditions, representing the weighting function of the Laplace operator, influences the localization properties of the eigenfunctions and the scaling of the integrated density of state (IDOS) N(lambda). The results indicate that N(lambda) deviates from the form given by the modified Weyl-Berry-Lapidus conjecture as it shows a correction of DeltaN(lambda) approximately lambda(D(f)/4) to the leading-order Weil term. Numerical results are presented for Koch and Koch snowflake fractal boundaries. The role of slip or no-slip boundary conditions of the velocity field on the IDOS is also investigated.


Pharmacokinetics of human leukocyte interferon.
Most of the human leukocyte interferon or rabbit serum interferon administered intravenously to rabbits was detectabble in their plasma 1 min after injection. An increase in the dose did not affect the early clearance of human interferon but did prolong its persistence in the serum. Repeated intramuscular injections had no effect on the kinetics of circulating interferon. The purity of the preparation influenced the rate at which interferon entered the blood after intramuscular or subcutaneous injection but did not affect clearance from the blood after intravenous injection. The circulating interferons obtained after intravenous or intramuscular administration cleared at similar rates. During the first 1-2 hr after intravenous or intramuscular injection into rats, the interferon levels were lower in lymph than in serum; after 2 hr lymph and serum samples contained similar amounts of interferon.


Electroretinography combined with spectral domain optical coherence tomography to detect retinal damage in shaken baby syndrome.
In order to correlate anatomical changes with visual function in shaken baby syndrome, we performed electroretinography and spectral domain optical coherence tomography on a 2-month-old girl and a 9-month-old girl after the retinal hemorrhages absorbed. Both patients had significant abnormalities in spectral domain optical coherence tomography images of the macular area. The amplitudes of the focal macular electroretinograms were more severely decreased than those of the full-field electroretinograms. Combining spectral domain coherence tomography with focal macular electroretinograms might better estimate the functional damage to the macula in patients with shaken baby syndrome.


Psychology ethics down under: a survey of student subject pools in Australia.
A survey of the 37 psychology departments offering courses accredited by the Australian Psychological Society yielded a 92% response rate. Sixty-eight percent of departments employed students as research subjects, with larger departments being more likely to do so. Most of these departments drew their student subject pools from introductory courses. Student research participation was strictly voluntary in 57% of these departments, whereas 43% of the departments have failed to comply with normally accepted ethical standards. It is of great concern that institutional ethics committees apparently continue to condone, or fail to act against, unethical research practices. Although these committees have a duty of care to all subjects, the final responsibility for conducting research in an ethical manner lies with the individual researcher.